计算溶液所需的质量、体积或浓度。
为什么选择此级别
，适用于对基线干扰要求严格的色谱和分析工作流程。
储存与运输
室温。常规运输 。请查阅批次 COA 获取详细规格。
质量文档
SDS、COA、产品数据表及规格说明书均可下载。可通过批号查询获取批次 COA。
文献证明
在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
概述
产品内容
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产品简介
本试剂盒适合于从新鲜唾液或唾液/保存液混合液中提取基因组DNA。本产品纯化过程不需使用苯酚或氯仿等有毒溶剂，无需乙醇沉淀。优化的缓冲体系使DNA异地结合到硅基质离心吸附柱上，PCR和其他酶促反应的抑制剂可通过两步洗涤步骤被有效去除，最后使用低盐缓冲液或水洗脱，即可得到高纯度DNA。纯化得到的可以直接用于酶切、PCR、Real-Time PCR、文库构建、Southern Blot、分子标记等下游实验。
自备试剂：无水乙醇
实验前准备及重要注意事项
1. 样品应避免反复冻融，否则会导致提取的DNA片段较小且提取量下降。
2. 第一次使用前应按照试剂瓶标签的说明在Buffer GW1和Buffer GW2中加入无水乙醇。
3. 使用前请检查Buffer GL是否出现结晶或者沉淀，如有结晶或者沉淀，请将Buffer GL于56℃水浴重新溶解。
4. 如果下游实验对RNA污染比较敏感，可以在第3步中加入4 μL DNase-Free的RNase A（100 mg/mL）。
5. 若要在室温下长时间保存唾液DNA。
操作步骤
1. 加入唾液样本或唾液/保存液混合液400μL 。
注意：1）加入保存液的唾液混合物需在提取前50℃水浴1小时或50℃空气温箱2小时。
2）如需要增加样本体积，则倍比增加步骤2-4中的Proteinase K、Buffer GL和无水乙醇的体积，步骤5中液体可分多次转入。
2. 加入40 μL Proteinase K。
3. 加入400 μL Buffer GL，涡旋震荡充分混匀，56℃水浴15-30分钟。
注意：如需去除RNA，可在上述步骤完成后，加入4μL浓度为100mg/mL的RNase A溶液，涡旋15秒，室温放置2分钟。
4. 短暂离心以去除管盖内壁的水珠。加入400μL无水乙醇，涡旋震荡充分混匀。短暂离心。
注意： 1) 加入Buffer GL和无水乙醇后要立即涡旋震荡混匀。
2) 加入Buffer GL和无水乙醇后可能会产生白色沉淀，不会影响后续实验。
3) GL和无水乙醇后可能形成溶胶状产物，此时推荐进行剧烈震荡或涡旋处理。
5. 上一个步骤中所得溶液全部加入到已装入收集管（Collection Tube）的吸附柱（Spin Column DM）中，若一次不能加完溶液，可分多次转入。12,000 rpm（～ 13,400 ×g）离心1分钟，倒掉收集管中的废液，将吸附柱重新放回收集管中。
6. 向吸附柱中加入500 μL Buffer GW1（使用前检查是否已加入无水乙醇），12,000 rpm 离心1分钟，倒掉收集管中的废液，将吸附柱重新放回收集管中。
7. 向吸附柱中加入500 μL Buffer GW2（使用前检查是否已加入无水乙醇），12,000 rpm 离心1分钟，倒掉收集管中的废液，将吸附柱重新放回收集管中。
注意：如需进一步提高DNA纯度，可重复步骤7。
8. 12,000 rpm离心2分钟，倒掉收集管中废液。将吸附柱置于室温数分钟，以彻底晾干。
注意：这一步的目的是将吸附柱中残余的乙醇去除，乙醇的残留会影响后续的酶促反应（酶切、PCR等）。
9. 将吸附柱置于一个新的离心管（自备）中，向吸附柱的中间部位悬空加入50-200 μL Buffer GE或灭菌水，室温放置2-5分钟，12,000 rpm离心1分钟，收集DNA溶液，-20℃保存DNA。
注意： 1）如果下游实验对pH值或EDTA敏感，可以用灭菌水洗脱。洗脱液的pH值对洗脱效率有很大影响，若用水做洗脱液应保证其pH值在7.0-8.5（可以用NaOH将水的pH值调到此范围），pH值低 于7.0时洗脱效率不高。
2） Buffer GE在65-70℃水浴预热，离心之前室温孵育5分钟可以增加产量。
3） 因为保存在水中的DNA会受到酸性水解作用的影响，如需长期保存，推荐用Buffer GE洗脱并于-20℃保存。
Product content
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Product Introduction
This kit is suitable for the extraction of genomic DNA from fresh saliva or saliva/preservation solution mixture.The purification process of this product does not require the use of toxic solvents such as phenol or chloroform, and ethanol precipitation is not necessary. The optimized buffer system enables DNA to bind heterogeneously to the silica matrix centrifugal adsorption column, and the inhibitors of PCR and other enzymatic reactions can be effectively removed by a two-step washing step, and finally eluted with a low-salt buffer or water to obtain high-purity DNA.The purified obtained can be directly used for enzyme digestion, PCR, Real-Time PCR, library construction, Southern Blot, molecular labeling and other downstream experiments.
Self-contained reagent: anhydrous ethanol
Pre-experiment Preparation and Important Notes
1. Repeated freezing and thawing of the sample should be avoided, as this may result in smaller fragments of extracted DNA and a decrease in the amount extracted.
2. Anhydrous ethanol should be added to Buffer GW1 and Buffer GW2 according to the instructions on the label of the reagent bottle before first use.
3. Before use, please check whether Buffer GL appears to be crystallized or precipitated.
Redissolve in a 56°C water bath.
4. If the downstream experiments are sensitive to RNA contamination, 4 μL DNase-Free RNase A can be added in step 3
(100 mg/mL).
5. For prolonged storage of salivary DNA at room temperature, our Salivary DNA Preservation Tubes are recommended.
Operation steps
1. Add 400 μL of saliva sample or saliva/preservation solution mixture.
Note: 1) Saliva mixtures added to the preservation solution require a 50°C water bath for 1 hour or an empty 50°C temperature chamber for 2 hours prior to extraction.
2) If an increase in sample volume is required, multiply the volumes of Proteinase K, Buffer GL, and anhydrous ethanol in Steps 2-4, and the liquid can be transferred in multiple times in Step 5.
2. Add 40 μL of Proteinase K.
3. Add 400μL Buffer GL, vortex and shake to mix thoroughly, and water bath at 56℃ for 15-30 minutes.
Note: If RNA removal is required, add 4 μL of RNase A solution at a concentration of 100 mg/mL after the above steps are completed, vortex for 15 seconds, and leave at room temperature for 2 minutes.
4. Centrifuge briefly to remove water droplets from the inside of the tube cap. Add 400 μL of anhydrous ethanol and mix well by vortexing and shaking. Centrifuge briefly.
Note: 1) Vortex and shake to mix immediately after adding Buffer GL and anhydrous ethanol.
The addition of Buffer GL and anhydrous ethanol may produce a white precipitate that will not affect subsequent experiments.
2) A sol-gel product may be formed after GL and anhydrous ethanol, in which case vigorous shaking or vortexing is recommended.
3) The solution obtained in the previous step is added to the adsorption column in the Collection Tube.
5. (Spin Column DM) in the collection tube, and if the solution cannot be added all at once, it can be transferred in several times. centrifuge at 12,000 rpm (∼13,400 × g) for 1 min, pour off the waste solution in the collection tube, and put the adsorption column back into the collection tube.
6. Add 500 μL of Buffer GW1 to the adsorption column (check that anhydrous ethanol has been added before use), centrifuge at 12,000 rpm for 1 minute, pour off the waste liquid in the collection tube, and put the adsorption column back into the collection tube.
7. Add 500 μL of Buffer GW2 to the adsorption column (check that anhydrous ethanol has been added before use), centrifuge at 12,000 rpm for 1 minute, pour off the waste liquid in the collection tube, and put the adsorption column back into the collection tube.
Note: Step 7 can be repeated if further DNA purity is required.
8. Centrifuge at 12,000 rpm for 2 minutes and pour off the waste liquid in the collection tube. Leave the adsorption column at room temperature for several minutes to dry thoroughly.
Note: The purpose of this step is to remove residual ethanol from the adsorption column, which can interfere with subsequent enzymatic reactions (digestion, PCR, etc.).
9. Place the adsorption column in a new centrifuge tube (supplied), add 50-200 μL of Buffer GE or sterilized water to the middle of the adsorption column overhanging the column, let it stand at room temperature for 2-5 minutes, and centrifuge at 12,000 rpm for 1 minute to collect the DNA solution.
-20°C to preserve DNA.
Note: 1) If the downstream experiment is sensitive to pH or EDTA, you can use sterilized water for elution. The pH of the eluent has a great influence on the elution efficiency, if water is used as the eluent should ensure that its pH is 7.0-8.5 (you can use NaOH to adjust the pH of the water to this range), and the elution efficiency is not high when the pH is lower than 7.0.
2) Buffer GE preheated in a 65-70°C water bath and incubated at room temperature for 5 min before centrifugation can increase the yield.
3) Because DNA preserved in water is subject to acidic hydrolysis, for long-term storage, elution with Buffer GE and storage at -20°C is recommended.
规格
别名
唾液基因组DNA提取试剂盒 | 唾液基因组DNA试剂盒
英文名称
Saliva Genomic DNA Kit
储存条件
室温
运输条件
常规运输
产品线
生物产品
名称和识别符
分子类型
试剂盒
技术文档
高级数据
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溶液计算器
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